LOCUS       sequence-418946-        7517 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cytosolic roGFP2-Orp1 (retroviral vector).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7517)
  AUTHORS   Gutscher M, Sobotta MC, Wabnitz GH, Ballikaya S, Meyer AJ, Samstag 
            Y, Dick TP
  TITLE     Proximity-based protein thiol oxidation by H2O2-scavenging 
            peroxidases.
  JOURNAL   J Biol Chem. 2009 Nov 13;284(46):31532-40. doi: 
            10.1074/jbc.M109.059246. Epub 2009 Sep 15.
  PUBMED    19755417
REFERENCE   2  (bases 1 to 7517)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1074/jbc.M109.059246"; journalName: "J Biol Chem"; date: 
            "2009-11-13- 13"; volume: "284"; issue: "46"; pages: "31532-40"
COMMENT     Sequence Label: pLPCX roGFP2-Orp1
FEATURES             Location/Qualifiers
     source          1..7517
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             472..647
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    710..909
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1110..1526
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1456..1478
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1494..1510
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     CDS             1623..2222
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1623..1642)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2119..2139
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     enhancer        2397..2700
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2701..2904
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2855..2875
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             join(2955..2957,2958..2960,2961..3671)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFISTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNCHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTCSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3000..3021)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3261..3280)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3608..3629
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     LTR             4314..4907
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5010..5032)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     protein_bind    5160..5193
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     protein_bind    5253..5286
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     5306..5325
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5406..5423)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5577..6165)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5657..5676)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6336..7127,7128..7196))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6959..6978
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7197..7301)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7369..7387
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tctaccctat tccccacact caccggagcc aaagccgcgg cccttccgtt tctttgcttt
       61 tgaaagaccc cacccgtagg tggcaagcta gcttaagtaa cgccactttg caaggcatgg
      121 aaaaatacat aactgagaat agaaaagttc agatcaaggt caggaacaaa gaaacagctg
      181 aataccaaac aggatatctg tggtaagcgg ttcctgcccc ggctcagggc caagaacaga
      241 tgagacagct gagtgatggg ccaaacagga tatctgtggt aagcagttcc tgccccggct
      301 cggggccaag aacagatggt ccccagatgc ggtccagccc tcagcagttt ctagtgaatc
      361 atcagatgtt tccagggtgc cccaaggacc tgaaaatgac cctgtacctt atttgaacta
      421 accaatcagt tcgcttctcg cttctgttcg cgcgcttccg ctctccgagc tcaataaaag
      481 agcccacaac ccctcactcg gcgcgccagt cttccgatag actgcgtcgc ccgggtaccc
      541 gtattcccaa taaagcctct tgctgtttgc atccgaatcg tggtctcgct gttccttggg
      601 agggtctcct ctgagtgatt gactacccac gacgggggtc tttcatttgg gggctcgtcc
      661 gggatttgga gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag
      721 caacttatct gtgtctgtcc gattgtctag tgtctatgtt tgatgttatg cgcctgcgtc
      781 tgtactagtt agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttct
      841 gaacacccgg ccgcaaccct gggagacgtc ccagggactt tgggggccgt ttttgtggcc
      901 cgacctgagg aagggagtcg atgtggaatc cgaccccgtc aggatatgtg gttctggtag
      961 gagacgagaa cctaaaacag ttcccgcctc cgtctgaatt tttgctttcg gtttggaacc
     1021 gaagccgcgc gtcttgtctg ctgcagcgct gcagcatcgt tctgtgttgt ctctgtctga
     1081 ctgtgtttct gtatttgtct gaaaattagg gccagactgt taccactccc ttaagtttga
     1141 ccttaggtca ctggaaagat gtcgagcgga tcgctcacaa ccagtcggta gatgtcaaga
     1201 agagacgttg ggttaccttc tgctctgcag aatggccaac ctttaacgtc ggatggccgc
     1261 gagacggcac ctttaaccga gacctcatca cccaggttaa gatcaaggtc ttttcacctg
     1321 gcccgcatgg acacccagac caggtcccct acatcgtgac ctgggaagcc ttggcttttg
     1381 acccccctcc ctgggtcaag ccctttgtac accctaagcc tccgcctcct cttcctccat
     1441 ccgccccgtc tctccccctt gaacctcctc gttcgacccc gcctcgatcc tccctttatc
     1501 cagccctcac tccttctcta ggcgccggaa tgatcccccg ggctgcaggt cggccgccac
     1561 gaccggtgcc gccaccatcc cctgacccac gcccctgacc cctcacaagg agacgacctt
     1621 ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc ccccgggccg
     1681 tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc gtcgacccgg
     1741 accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc gtcgggctcg
     1801 acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg accacgccgg
     1861 agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc gagttgagcg
     1921 gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac cggcccaagg
     1981 agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc aagggtctgg
     2041 gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg cccgccttcc
     2101 tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc accgtcaccg
     2161 ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag cccggtgcct
     2221 gacgcccgcc ccacgacccg cagcgcccga ccgaaaggag cgcacgaccc catggctccg
     2281 accgaagcca cccggggcgg ccccgccgac cccgcacccg cccccgaggc ccaccgactc
     2341 tagtaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     2401 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     2461 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     2521 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     2581 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     2641 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     2701 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
     2761 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
     2821 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
     2881 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
     2941 actcagatct cgagatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     3001 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     3061 gcgatgccac ctacggcaag ctgaccctga agttcatctc caccaccggc aagctgcccg
     3121 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     3181 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     3241 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     3301 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     3361 acatcctggg gcacaagctg gagtacaact acaactgcca caacgtctat atcatggccg
     3421 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     3481 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     3541 tgcccgacaa ccactacctg agcacctgct ccgccctgag caaagacccc aacgagaagc
     3601 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     3661 agctgtacaa gactagtggt ggttcaggtg gtggtggttc aggtggtggt ggttcaggtg
     3721 gaggaggatc aggaggagga ggatcaggag gaggaggatc aggaggagaa ttcgatatct
     3781 cagaattcta taagctagca cctgttgaca agaaaggcca accattcccc ttcgaccaat
     3841 taaagggaaa agtggtgctt atcgttaatg ttgcctccaa atgtggattc actcctcaat
     3901 acaaagaact agaggccttg tacaaacgtt ataaggacga aggatttacc atcatcgggt
     3961 tcccatgcaa ccagtttggc caccaagaac ctggctctga tgaagaaatt gcccagttct
     4021 gccaactgaa ctatggcgtg actttcccca ttatgaaaaa aattgacgtt aatggtggca
     4081 atgaggaccc tgtttacaag tttttgaaga gccaaaaatc cggtatgttg ggcttgagag
     4141 gtatcaaatg gaattttgaa aaattcttag tcgataaaaa gggtaaagtg tacgaaagat
     4201 actcttcact aaccaaacct tcttcgttgt ccgaaaccat cgaagaactt ttgaaagagg
     4261 tggaatagat cgataaaata aaagatttta tttagtctcc agaaaaaggg gggaatgaaa
     4321 gaccccacct gtaggtttgg caagctagct taagtaacgc cattttgcaa ggcatggaaa
     4381 aatacataac tgagaataga gaagttcaga tcaaggtcag gaacagatgg aacagctgaa
     4441 tatgggccaa acaggatatc tgtggtaagc agttcctgcc ccggctcagg gccaagaaca
     4501 gatggaacag ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc
     4561 tcagggccaa gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac
     4621 catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta
     4681 accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag
     4741 agcccacaac ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc
     4801 gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga
     4861 gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc
     4921 gggatcggga gacccctgcc cagggaccac cgacccacca ccgggaggta agctggctgc
     4981 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
     5041 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     5101 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtactga
     5161 taacttcgta taatgtatgc tatacgaagt tattaggtct gaagaggagt ttacgtccag
     5221 ccaagctagg gccgcgatcc ggaaccctta atataacttc gtataatgta tgctatacga
     5281 agttatcagt actggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat
     5341 atgcggtgtg aaataccgca cagatgcgta aggagaaaat accgcatcag gcgctcttcc
     5401 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     5461 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     5521 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     5581 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     5641 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5701 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5761 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5821 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5881 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5941 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     6001 tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     6061 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     6121 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     6181 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     6241 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     6301 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     6361 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     6421 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     6481 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     6541 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     6601 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tgcaggcatc
     6661 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6721 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6781 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6841 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6901 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     6961 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     7021 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     7081 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     7141 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     7201 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     7261 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     7321 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     7381 acgaggccct ttcgtcttca agaattgcta gcaattgcta gcaattgcta gcaattcata
     7441 ccagatcacc gaaaactgtc ctccaaatgt gtccccctca cactcccaaa ttcgcgggct
     7501 tctgcctctt agaccac
//
